title: miR-335 and miR-363 regulation of neuroblastoma tumorigenesis and metastasis.
abstract: BACKGROUND
  microRNA (miRNA) functions broadly as post-transcriptional regulators of gene expression, and disproportionate miRNAs can result in dysregulation of oncogenes in cancer cells. We have previously shown that gastrin-releasing peptide receptor (GRP-R) signaling regulates tumorigenicity of neuroblastoma cells. Herein, we sought to characterize miRNA profile in GRP-R silenced neuroblastoma cells, and to determine the role of miRNAs on tumorigenicity and metastatic potential.
  
  
  METHODS
  Human neuroblastoma cell lines, BE(2)-C and SK-N-SH, were used for our study. Stably transfected GRP-R silenced cells were assessed for miRNA profiles. Cells were transfected with miR-335, miR-363, or miR-CON, a nontargeting control, and in vitro assays were performed. In vivo functions of miR-335 and miR-363 were also assessed in a spleen-liver metastasis murine model.
  
  
  RESULTS
  GRP-R silencing significantly increased expression of miR-335 and miR-363 in BE(2)-C cells. Overexpression of miR-335 and miR-363 decreased tumorigenicity as measured by clonogenicity, anchorage-independent growth, and metastasis determined by cell invasion assay and liver metastasis in vivo.
  
  
  CONCLUSIONS
  We report, for the first time, that GRP-R-mediated tumorigenicity and increased metastatic potential in neuroblastoma are regulated, in part, by miR-335 and miR-363. A better understanding of the anti-tumor functions of miRNAs could provide valuable insights to discerning molecular mechanisms responsible for neuroblastoma metastasis.
authors.affiliation: Department of Pediatric Surgery, Vanderbilt University Medical Center, Nashville, TN, USA.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Qiao
authors.forename: Sora
authors.initials: S
authors.lastname: Lee
authors.forename: Pritha
authors.initials: P
authors.lastname: Paul
authors.forename: Lauren
authors.initials: L
authors.lastname: Theiss
authors.forename: Joshua
authors.initials: J
authors.lastname: Tiao
authors.forename: Lan
authors.initials: L
authors.lastname: Qiao
authors.forename: Andrew
authors.initials: A
authors.lastname: Kong
authors.forename: Dai H
authors.initials: DH
authors.lastname: Chung
mesh_terms: D051722:ADAM Proteins / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D006801:Humans; D008114:Liver Neoplasms, Experimental / Q000556:secondary; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000502:physiology*; D009361:Neoplasm Invasiveness; D009447:Neuroblastoma / Q000209:etiology* / Q000473:pathology / Q000556:secondary; D018004:Receptors, Bombesin / Q000032:analysis / Q000502:physiology
chemical_list: C556347:MIRN335 microRNA, human; C574861:MIRN363 microRNA, human; D008565:Membrane Proteins; D035683:MicroRNAs; D018004:Receptors, Bombesin; D051722:ADAM Proteins; C098660:ADAM15 protein, human